SAGA deletion Module

General notes

Handouts

Daily Notes

Notes:M3D1

need forward and reverse primers for all non-essential SAGA subunits ordered, validated and available

need template (pRS406) available

need 2.5X PCR "master mix" made up

have students open Primer Record Handout to use as a guide in primer design

Materials

Forward primer

Reverse primer

Template – 40ul/team

Pcr master mix – 1ul/team

Notes:M3D2

need quiz for this day to start lab

need FY2068 ON then subcultured flask growing for lab

need YPD plates, and -ura and a few -trp (2 YPD, 2 -ura, 2 -trp per lab group for viability assay)

aliquots for each pair of students: 1 ml ON culture, 1 ml of log phase for counting and viability assays, additional 10 ml aliquot of log phase for prep of competent cells, bottles of sterile H20 (students can keep these in their drawers if that's easier), cuvettes